Anti-SHP1 antibody [Y476]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Tissue Microarrays stained for "Anti-SHP1 antibody [Y476]" using "ab32559"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab32559 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Unpurified ab32559, at a 1/50 dilution, staining human lymph node by immunohistochemistry, Paraffin embedded tissue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Immunohistochemical staining of paraffin embedded human tonsil with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Tissue Microarrays stained for "Anti-SHP1 antibody [Y476]" using "ab32559"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab32559 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Unknown

Western blot - Anti-SHP1 antibody [Y476] (AB32559)

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution

Lane 1:

A- A431 cell lysate

Lane 2:

B- Jurkat cell lysate

Lane 3:

C- K562 cell lysate

Predicted band size: 68 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-SHP1 antibody [Y476] (AB32559)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution

Lane 1:

A431 cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 68 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-SHP1 antibody [Y476] (AB32559)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution

Lane 1:

SP2/0 cell lysate at 20 µg

Lane 2:

mouse marrow at 20 µg

Lane 3:

rat brain at 20 µg

Lane 4:

C6 cell lysate at 20 µg

Lane 5:

rat cerebral cortex at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 68 kDa

Observed band size: 65 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SHP1 antibody [Y476] (AB32559)

Immunohistochemical staining of paraffin embedded rat spleen with purified ab32559 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Lab

Western blot - Anti-SHP1 antibody [Y476] (AB32559)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32559 observed at 70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab32559 was shown to react with SHP1 in wild-type THP-1 cells in Western blot with loss of signal observed in PTPN6 knockout sample. Wild-type THP-1 and PTPN6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab32559 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SHP1 antibody [Y476] (ab32559) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

PTPN6 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN6 knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-ptpn6-knockout-thp-1-cell-line-ab273717'>ab273717</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 68 kDa,83 kDa

Observed band size: 70 kDa,85 kDa

false